Molecular characterization and xenogenic application of wharton’s jelly derived caprine mesenchymal stem cells

Aim of the present study was in vitro expansion and characterization of caprine wharton’s jelly derived mesenchymal stem cells (cWJ-MSCs) to investigate their tissue healing potential in xenogenic animal model. Plastic adherent fibroblastoid cell populations with distinctive homogeneous morphology were isolated from caprine wharton’s jelly explants. These wharton’s jelly derived cells were found positive for the surface markers CD-73, STRO-1 and CD-105, whereas they were negative for hematopoetic stem cell marker CD-34. In vitro cultured cWJ-MSCs also showed differentiation properties into osteogenic, adipogenic and chondrogenic lineages as demonstrated by von Kossa, Oil Red- O and Alcian blue staining respectively, which was further confirmed and quantified by flow cytometric analysis. Furthermore, these well characterized cWJ-MSCs were evaluated for the wound-healing potential in full-thickness skin wounds in rabbit model for 28 days. Caprine WJ- MSCs treated skin wounds showed significantly (P?<?0.05) higher percentage of wound contraction especially at the 21^st day post transplantation when compared to PBS treated control group animals. Further, we observed better healing potential of cWJ-MSCs in terms of histo-morphological evaluation, epithelialisation and collagenization with matured vascularization stage by day 28 as compared to control. In conclusion, cWJ- MSCs provide an alternative inexhaustible source of mesenchymal stem cells and also unravel new perspectives pertaining to the therapeutic use of these cells in different species.     

Porcine parvovirus- and porcine circovirus 2-associated reproductive failure and neonatal mortality in crossbred Indian pigs

The objective of this study was to detect the presence of porcine parvovirus (PPV) and porcine circovirus 2 (PCV2) in a farm showing reproductive failure and increased mortality in neonatal piglets by histopathological examination, polymerase chain reaction, and demonstration of viral antigen and nucleic acid. Out of 594 piglets farrowed by 70 first-parity gilts, nine (1.51%) mummified fetuses, 13 (2.19%) stillborn, and 572 (96.3%) live-born piglets were recorded. The average litter size at birth was 8.48 piglets per litter. One hundred ninety-four (33.91%) piglets died within 7 days of age. PPV was detected in five litters (7.14%) and two of them revealed coinfection with PCV2. The pathological lesions in the coinfected litters were more severe, indicating a synergistic action between the two viruses. Results of this study suggest for the first time occurrence of PPV and coinfection with PCV2 in crossbred Indian pigs affected with reproductive problem and neonatal mortality.     

Molecular and Cellular Characterization of Buffalo Bone Marrow‐Derived Mesenchymal Stem Cells

Immune privileged mesenchymal stem cells (MSCs) can differentiate into multiple cell types and possess great potential for human and veterinary regenerative therapies. This study was designed with an objective to isolate, expand and characterize buffalo bone marrow‐derived MSCs (BM‐MSCs) at molecular and cellular level. Buffalo BM‐MSCs were isolated by Ficoll density gradient method and cultured in Dulbecco’s modified Eagle’s medium supplemented with fetal bovine serum (FBS). These cells were characterized through alkaline phosphatase (AP) staining, colony‐forming unit (CFU) assay, mRNA expression analysis (CD 73, CD 90, CD 105, Oct4 and Nanog), immunolocalization along with flow cytometry (Stro 1, CD 73, CD 105, Oct4, Sox2 and Nanog) and in situ hybridization (Oct4 and Sox2). Multilineage differentiation (osteogenic, adipogenic and chondrogenic) was induced in vitro, which was further assessed by specific staining. Buffalo BM‐MSCs have the capacity to form plastic adherent clusters of fibroblast‐like cells and were successfully maintained up to 16^th passage. These cells were AP positive, and further CFU assay confirmed their clonogenic property. RT‐PCR analysis and protein localization study showed that buffalo BM‐MSCs are positive for various cell surface markers and pluripotency markers. Cytoplasmic distribution of mRNA for pluripotency markers in buffalo BM‐MSCs and multilineage differentiation were induced in vitro, which was further assessed by specific staining. To the best of our knowledge, this is the first report of buffalo BM‐MSCs, which suggests that MSCs can be derived and expanded from buffalo bone marrow and can be used after characterization as a novel agent for regenerative therapy.     

Marker‐assisted breeding of Pi‐1 and Piz‐5 genes imparting resistance to rice blast in PRR78, restorer line of Pusa RH‐10 Basmati rice hybrid

Rice blast, caused by fungus Magnaporthe grisea, is a serious disease causing considerable economic damage worldwide. Best way to overcome disease is to breed for disease‐resistant cultivars/parental lines of hybrids. Pusa RH10, first aromatic, fine‐grain rice hybrid released and cultivated extensively in India. Hybrid and its parental lines, Pusa 6A and PRR78, are highly susceptible to blast. CO39 pyramid carrying two dominant, broad‐spectrum blast‐resistance genes, viz. Pi‐1 and Piz‐5, used as a donor parent to introgress these genes into PRR78 using marker‐assisted backcrossing (MABC). Microsatellite markers RM5926 and AP5659‐5 tightly linked to Pi‐1 and Piz‐5 genes, respectively, were used for foreground selection to derive introgression lines. Further, these lines were evaluated for agronomic performance, disease reaction and cooking quality traits along with PRR78. Most of the improved lines were on par with PRR78 for all traits evaluated except gelatinization temperature. Recurrent parent genome percentage (RPG) study also revealed similarity of these lines with PRR78. Hybrids derived using improved PRR78 lines were superior over Pusa RH10 in terms of yield.     

Expression and Characterization of Constitutive Heat Shock Protein 70.1 (HSPA‐1A) Gene in In Vitro Produced and In Vivo‐Derived Buffalo (Bubalus bubalis) Embryos

Cells are blessed with a group of stress protector molecules known as heat shock proteins (HSPs), amongst them HSP70, encoded by HSPA‐1A gene, is most abundant and highly conserved protein. Variety of stresses hampers the developmental competence of embryos under in vivo and in vitro conditions. Present work was designed to study the quantitative expression of HSPA‐1A mRNA in immature oocytes (IMO), matured oocytes (MO), in vitro produced (IVP) and in vivo‐derived (IVD) buffalo embryos to assess the level of stress to which embryos are exposed under in vivo and in vitro culture conditions. Further, HSPA‐1A gene sequence was analysed to determine its homology with other mammalian sequences. The mRNA expression analysis was carried out on 72 oocytes (40 IMO; 32 MO), 76 IVP and 55 IVD buffalo embryos. Expression of HSPA‐1A was found in oocytes and throughout the developmental stages of embryos examined irrespective of the embryo source; however, higher (p < 0.05) expression was observed in 8–16 cell, morula and blastocyst stages of IVP embryos as compared to IVD embryos. Phylogenetic analysis of bubaline HSPA‐1A revealed that it shares 91–98% identity with other mammalian sequences. It can be concluded that higher level of HSPA‐1A mRNA in IVP embryos in comparison with in vivo‐derived embryos is an indicator of cellular stress in IVP system. This study suggests need for further optimization of in vitro culture system in which HSPA‐1A gene could be used as a stress biomarker during pre‐implantation development.     

Transgressive segregation for yield traits in Oryza sativa IR58025B X Oryza meridionalis Ng. Bc2F3population under irrigated and aerobic conditions

Wild species of the genus Oryza are a good source of beneficial alleles for enhancing rice yield under normal and adverse conditions. BC₂F₃ population was derived from a cross between Oryza sativa IR58025B and Oryza meridionalis Ng. (2n = 24, AA) a heat tolerant wild species to evaluate 12 yield traits under irrigated and aerobic conditions. Analysis of variance and genetic estimates indicated there is substantial genetic variation among progenies under both conditions. Grain yield had high heritability (61.9%) and genetic advance (36.4%) under irrigated conditions but moderate heritability (49.6%) and genetic advance (13.3%) under aerobic conditions indicating that selection for yield will be effective under both conditions. Panicle number, grain number, spikelet fertility, and test weight showed significant positive correlation with grain yield under both conditions. Families out-performing IR58025B for yield under both conditions were obtained providing evidence that phenotypically inferior O. meridionalis contributed to yield increase. This species can be a novel source of natural genetic variation for the improvement of rice under irrigated as well as under aerobic condition.     

Identification and genotypic characterization of porcine teschovirus from selected pig populations in India

Tropical Animal Health and Production - Porcine teschovirus (PTV) previously classified as porcine enteroviruses in the family Picornaviridae are associated with a wide range of illnesses in swine...